The Culture Of DM-derived Adipose Stem Cells And The Differentiation Into Endothelial Cells In Vitro

BackgroundAs a long-term global threat to human health problems, diabetes s chronic complication often leads to multiple system organ dysfunction and failure, but the current treatment methods are not ideal. In the chronic complications of diabetes, vascular lesion is the main pathogenic factors, and the key target tissue of it is endothelial cells. Endothelial cells has played an important role in regulating vascular tension, vascular wall s permeability and involved in angiogenesis, intracellular signal transduction. However, endothelial cells perform dysfunction in disease, which makes it become a promising treatment by transplanting endothelial cells or stem cells. As a excellence seed cell of vascular tissue engineering, ADSCs not only exhibit prominent differentiation potency, but also have many advantages, such as wide source, easy access, quickly proliferation, minimally invasion. At present many studies have confirmed the capacity about ADSC differentiating into endothelial cells, but the research of DM-derived ADSC is few. For the realization of autologous DM-derived ADSC transplantation, it is a key point whether DM-derived ADSC hold the same character with normal ADSC.Objective1. To found the process of culturing and identifying DM-derived ADSC, and comparing with normal ADSC.2. To study the capacity of DM-derived ADSC differentiating into endothelial cell, providing theory evidence for autologous DM-derived ADSC transplantation.MethodExperimental group:DM-derived ADSCControl group:normal ADSCTaking adipose tissue from normal person and DM patient, and isolating and culturing two group cells by the process of normal ADSC; Taking P3generation ADSC, and identifying the cell markers of two group cells, including CD29, CD44, CD105, by using flow cytometer; Taking P3generation ADSC, and inducing them to differentiate into endothelial cells in24well plates by using ES-inducing medium; observing and comparing the morphological change of two group cells during the inducing period, and taking6d,9d,12d s cells after inducing to process immunocytochemistry staining of CD31.ResultThe typical ADSC could be achieved from DM s adipose tissue by the similar method of normal ADSC isolation and culture; flow cytometer identified CD29(+), CD44(+), CD105(+) on P3generation cells coming from both two groups; the two group cells both changed modality after being induced six days and appeared to CD31(+) by immunocytochemistry staining after being induced nine days; after twelve days, the typical modality of endothelial cells could be observed.Conclusion1. Found the preliminary method of isolating and culturing DM-derived ADSC;2. Prove the self-renewal capacity of DM-derived ADSC, which could be compared with normal ADSC.3. Prove the differentiation capacity to endothelial cells of DM-derived ADSC...